GSK-3 inhibitor 1_603272-51-1_DataSheet_MedChemExpress

SGLT2抑制剂联合二甲双胍治疗对糖尿病肾病患者血糖及疗效的影响郑秋娥，程秋敏，刘江建福建省立医院药学部，福建福州350001[摘要]目的分析钠-葡萄糖共转运蛋白2（sodium-dependent glucose transporters 2, SGLT2）抑制剂联合二甲双胍治疗对糖尿病肾病患者血糖及疗效的影响。

方法选取2021年1月—2023年1月福建省立医院接诊的108例糖尿病肾病患者，按照随机数表法分为对照组与研究组，各54例。

对照组接受二甲双胍+百令胶囊治疗，研究组联合SGLT2抑制剂治疗，分析对比两组患者血糖指标、临床总有效率、肾功能指标及血清炎症指标。

结果与对照组相比，研究组治疗后的空腹血糖、餐后2 h血糖及糖化血红蛋白水平更低，差异有统计学意义（P<0.05）。

与对照组对比，研究组临床总有效率更高，差异有统计学意义（P<0.05）。

与对照组对比，研究组治疗后的血肌酐、尿素氮及尿白蛋白排泄率更低，差异有统计学意义（P<0.05）。

结论SGLT2抑制剂联合二甲双胍治疗糖尿病肾病可降低血糖水平，改善肾功能，减轻机体炎症反应，提高临床总效率。

[关键词] SGLT2抑制剂；二甲双胍；糖尿病肾病；血糖[中图分类号] R4 [文献标识码] A [文章编号] 1672-4062（2023）10（b）-0076-04Effect of SGLT2 Inhibitor Combined with Metformin Treatment on Blood Glucose and Curative Effect in Patients with Diabetic NephropathyZHENG Qiu'e, CHENG Qiumin, LIU JiangjianDepartment of Pharmacy, Fujian Provincial Hospital, Fuzhou, Fujian Province, 350001 China[Abstract] Objective To analyze the effects of sodium-dependent glucose transporters 2 (SGLT2) inhibitor combined with metformin treatment on blood glucose and curative effect in patients with diabetic nephropathy.Methods 108 pa⁃tients with diabetes nephropathy who were treated in Fujian Provincial Hospital from January 2021 to January 2023 were selected and divided into the control group and the study group according to the random number table, with 54 patients in each group. The control group received treatment with metformin and Bailing capsules, while the study group received treatment with SGLT2 inhibitors. The blood glucose indicators, clinical total efficiency, renal function indicators, and serum inflammation indicators were analyzed and compared between the two groups.Results Compared with the control group, the study group had lower levels of FPG, 2 hPG, and HbA1c after treatment, the difference was statistically significant (P<0.05). Compared with the control group, the total clinical efficiency of the study group was higher, and the difference was statistically significant (P<0.05). Compared with the control group, the study group had lower Scr, BUN, and UAER after treatment, the difference was statistically significant (P<0.05).Conclusion SGLT2 in⁃hibitor combined with metformin in the treatment of diabetic nephropathy can reduce blood glucose level, improve re⁃nal function, reduce the inflammatory response of the body, and improve the overall clinical efficiency.[Key words] SGLT2 inhibitor; Metformin; Diabetic nephropathy; Blood glucose糖尿病肾病是糖尿病并发症之一，也是引发终末期肾衰竭的主要原因，近十年来随着糖尿病发病率的增高，糖尿病肾病发病率增加2倍以上[1]。

Sutent药物基本信息〖NDA申请人〗CPPY CV〖NDA原始批准日期〗2006年07月26日〖剂型/规格〗胶囊剂/12.5mg；胶囊剂/25mg；胶囊剂/50mg；胶囊剂/37.5mg〖适应证〗50mg QD，用于治疗：Ⅰ、病情恶化后或对马来酸伊马替尼不耐受的胃肠间质瘤；Ⅱ、晚期肾细胞瘤活性成分信息〖USAN名称〗Sunitinib Malate，苹果酸舒尼替尼〖CAS号〗341031-54-7（苹果酸盐）；557795-19-4（游离碱）〖曾用代号〗SU-11248（苹果酸盐）〖作用类别〗激酶抑制剂类抗肿瘤药〖化学名〗(Z)-N-(2-(二乙基氨基)乙基)-5-((5-氟-2-氧代吲哚-3-亚基)甲基)-2,4-二甲基-1H-吡咯-3-羧酰胺苹果酸盐〖化学结构式〗专利信息年度销售情况（亿美元，信息来源：辉瑞公司年度财务报告及SEC报表）Tykerb药物基本信息〖NDA申请人〗Smithkline Beecham〖NDA原始批准日期〗2007年03月13日〖剂型/规格〗片剂/250mg；〖适应证〗1250mg QD+卡培他滨治疗肿瘤过度表达HER2且使用过包括蒽环类抗生素、紫杉烷类抗生素曲妥珠单抗在内的抗肿瘤药物治疗的晚期或转移性乳腺癌；1500 QD+来曲唑治疗HER2过度表达且需要进行激素治疗的绝经后妇女的激素受体阳性的转移性乳腺癌活性成分信息〖USAN名称〗Lapatinib ditosylate （monohydrate），拉帕替尼二（对甲基苯磺酸）盐（单水合物）〖CAS号〗388082-78-8〖曾用代号〗〖作用类别〗激酶抑制剂类抗肿瘤药；〖化学名〗N-[3-氯-4-[(3-氟苯基)甲氧基]苯基]-6-[5[[[2-(甲磺酰基)乙基]氨基]甲基]-2-呋喃基]-4-喹啉胺二（对甲基苯磺酸）盐单水合物〖理化性质〗黄色固体，25℃下于水中的溶解度为0.007mg/mL，于0.1N HCl中的溶解度为0.001mg/mL〖化学结构式〗专利信息年度销售情况（亿英磅）Tasigna药物基本信息〖NDA申请人〗诺华制药〖NDA原始批准日期〗2007.10.29〖剂型/规格〗片剂/200mg（按游离碱计）〖适应证〗300mg BID用于于慢性期治疗新近确认成年患者的费城染色体阳性慢性髓样白血病；400mg BID用于于慢性期或急性期治疗成年患者对包括伊马替尼在内的先前治疗方法耐药或不耐受的费城染色体阳性慢性髓样白血病。

Effect of GSK-3Inhibitor SB216763on Expression of Glycogen synthasekinese-3and Insulin Receptor Substrates-1of Granulosa cellsin PCOS.AbstractObjective:Polycystic ovarian syndrome(PCOS)is a common reproductive disorder, and its pathogenesis has Insulin resistance(IR).To investigate the effects of GSK-3 inhibitor SB216763on expression of Glycogen synthase kinese-3β(GSK-3β)and Insulin Receptor Substrates-1(IRS-1)in Granulosa cells of PCOS.Methods:Ovarian granulosa cells(GC)were collected from PCOS patients undergoing in vitro fertilization/intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET)(n=20).Every sample of GC was inoculated in two culture bottles and cultured for3days.GSK-3inhibitor was added in one of them,and thus the two bottle of GC were divided into PCOS+IN group and PCOS group.Those ovarian GC from non-PCOS patients undergoing IVF/ICSI-ET used as control group(N-PCOS group)(n=16).The cultured ovarian GC of three groups were examined by Western Blotting for GSK-3,phosphorylated of p-GSK-3β(Ser9),and of p-IRS-1(Ser312).Result:The protein expression levels of GSK-3、p-GSK-3βin the GC of PCOS group were lower than N-PCOS group,while p-IRS-1were higher than N-PCOS group, statistically significant between the groups(P＜0.05).The protein expression levels of p-GSK-3βafter SB216763treatment(PCOS+IN)were lower than N-PCOS group,but were higher than PCOS group,mean while p-IRS-1were higher than N-PCOS group,but were lower than PCOS group,statistically significant between the groups(P＜0.05).The difference of protein expression level of GSK-3between PCOS and PCOS+IN group was not significant(P＞0.05).Conclusion:Activity of AKT/GSK-3signaling pathway in ovarian GC of PCOS is higher than non-PCOS.There is a link between IRS-1/MAPK signaling pathway and AKT/GSK-3signaling pathway,SB216763may decrease the activity of GSK-3and the expression of p-IRS-1.Postgraduate student:Yuan-yuan Wang（Obstetrics and Gynecology）Directed by Prof:Jian-xin LiuKey Words:Polycystic ovarian syndrome(PCOS);Granulosa cell(GC);Inhibitor; GSK-3；IRS-1目录引言 (1)材料和方法 (4)1研究对象 (4)2临床治疗方案 (4)3实验仪器与试剂 (5)3.1主要仪器 (5)3.2主要试剂及耗材 (6)4实验方法 (7)4.1主要溶液配置方法 (7)4.2黄素化颗粒细胞的提纯、原代培养及镜下观察 (9)4.3Western Blot检测各组GSK-3、p-GSK-3β（Ser9）、p-IRS-1（Ser312）表达量 (9)5统计学方法 (11)结果 (12)1患者一般资料 (12)2黄素化颗粒细胞的原代培养情况 (12)3Westen Blot检测各组颗粒细胞蛋白表达情况 (13)讨论 (14)1PCOS患者卵巢局部胰岛素作用途径缺陷 (15)2GSK-3抑制剂对PCOS卵巢局部IR作用 (16)3问题与展望 (17)结论 (18)参考文献 (19)综述 (22)综述参考文献 (31)附录 (36)攻读学位期间的研究成果 (38)致谢 (39)学位论文独创性声明、学位论文知识产权权属声明 (40)引言引言多囊卵巢综合征（Polycystic ovary syndrome，PCOS）作为一种妇科常见的生殖内分泌紊乱性疾患，临床患者主要表现为机体糖代谢异常及卵巢生殖功能障碍。

GSK3β、p-GSK3βser9、p-GSK3βTyr216在宫颈癌组织中的表达及临床意义陈良凤;张颖;王仲奇;邸曼;郑福利;王建【摘要】Objective To investigate the expressions of glycogen synthase kinase 3β( GSK3β) , p-GSK3βser9 and p-GSK3βTyr216 in cervical carcinoma and their correlation with clinical pathological factors.Methods By using immunohistochemistry, GSK3β, p-GSK3βser9 and p-GSK3βTyr216 protein expressions were detected in 80 cases of cervical carcinoma admitted in Xijing Hospital of Fourth Military Medical University during January 2012 to December 2014.The correlations between expressions of GSK3β, p-GSK3βser9 and p-GSK3βTyr216 and clinicopathological factors were analyzed.Results The positive expression rates of GSK3β, p-GSK3βser9 and p-GSK3βTyr216 in cervical carcinoma were 18.8%, 68.8% and 12.5%,resp ectively.The expressions of GSK3βand p-GSK3βTyr216 in early stage group were significantly higher than those in advanced stage group (25.5%vs 4.0%, χ2 =5.193,P=0.029;76.4% vs 52.0%,χ2 =4.749,P=0.039), but the expression rate of p-GSK3βser9 was obviously lower (5.5%vs 28.0%,χ2 =7.988,P=0.009).The positive expression rates of p-GSK3βser9 in well and moderately differentiated tumors were lower than those in poorly differentiated tumors (7.4%, 7.9%and 33.3%,χ2 =7.329, P=0.031).The expressions of GSK3βand p-GSK3βTyr216 were significantly lower in patients with positive lymph node metastasis than in those with negative lymph node status (5.9%vs 28.3%,χ2 =6.427,P=0.018;52.9%vs 80.4%,χ2=6.878, P=0.014), but the expression of p-GSK3βser9 was higher in patients with lymph node metastases (23.5% vs 4.3%, χ2=6.577,P=0.015) .Conclusion The expressions of GSK3β, p-GSK3βser9 andp-GSK3βTyr216 in cervical carcinoma are correlated with clinical TNM staging, histological grades and lymph node metastases, which suggests that GSK3βmay play a potential role in cervical tumor genesis.%目的：检测糖原合成酶激酶3β（GSK3β）、p-GSK3βser9和p-GSK3βTyr216表达水平在宫颈癌组织中的表达情况及其与临床病理因素的相关性。

中国兽医科学2021,51(02〉： 161-168Chinese Veterinary Science网络首发时间：2020-12-ll D01：10.16656/j.issn.l673-4696.2021.0024 中图分类号：S852.734 文献标志码：A文章编号：1673_4696(2021)02-016卜08猪带纟条虫丝氨酸蛋白酶抑制剂Ts-serpin-1对 人巨噬细胞THP-1的免疫调节功能研究毕研丽\刘仲蓉'郭爱疆\张少华、王帅”，才学鹏(1.中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室，甘肃兰州730046;2.甘肃农业大学动物医学院，甘肃兰州730070;3.中国兽医药品监察所，北京100081)摘要：主要研究猪带绦虫丝氨酸蛋白酶抑制剂Ts-serpin-l(WormBase:TsM_000065700)对宿主THP-1 细胞的免疫调节作用通过设计特异性引物和RT-PCR扩增技术，获得Ts-serpin-1编码序列，用qRT-PCR 分析Ts-serpin-l基因在猜带線虫成虫和中綠期幼虫的表达情况；构建pCold-Ts-serpin-1原核表达栽体，诱导表达纯化重组蛋白Ts-serpin-1;用重组蛋白Ts-serpin-1处理THP-1细胞，采用qRT-PCR和ELISA方法检测Ts-serpin-1处理THP-1细胞后，各炎性细胞因子的变化情况，结果显示：获得的Ts-serpin-1目的基因长度为1149匕口，编码382个氨基酸，含有56卬丨11家族特有的反应中心环。

7^-36叩丨11-1基因在猪带绦虫 成虫和中绦期幼虫均表达，且成虫表达量显著高于幼虫。

重组蛋白Ts-serpin-1的分子质量约为43 ku,可抑 制THP-1细胞促炎性细胞因子IL-6、IL-10、IL- 12、TNF-a、丨FN-y和iNOS2的表达，促进抗炎性细胞因子 1L-10和TGF-y3的分泌表达。

网络出版时间：２０２４－０３－０９１８：５９：３０　网络出版地址：ｈｔｔｐｓ：／／ｌｉｎｋ．ｃｎｋｉ．ｎｅｔ／ｕｒｌｉｄ／３４．１０８６．Ｒ．２０２４０３０６．１７２８．０５４◇实验方法学◇基于实时无标记细胞分析技术时间维度特征的体外抗肿瘤活性评价新策略刘芳彤１，邢淑雁２，刘孝云３，４，５，６，叶冬雪３，４，５，６，杨　佳２，张国英２，容　蓉２，４，杨　勇３，４，５，６（山东中医药大学１．中医药创新研究院、２．药学院、３．实验中心，４．中医药经典理论教育部重点实验室，５．中医药基础研究山东省重点实验室，６．山东省中医药抗病毒工程研究中心，山东济南　２５０３５５）收稿日期：２０２３－１２－０５，修回时间：２０２４－０１－１５基金项目：国家自然基金资助项目（Ｎｏ２１８０７０６６）作者简介：刘芳彤（１９９７－），女，硕士生，研究方向：中医药抗病毒的药效及机制，中医药抗肿瘤的生物疗法探索，Ｅ ｍａｉｌ：ｌｉ ｕｆａｎｇｔｏｎｇ２０２１＠１６３．ｃｏｍ；容　蓉（１９７０－），女，博士，教授，研究方向：中药及复方活性成分与质量控制，通信作者，Ｅ ｍａｉｌ：ｒｏｓｉｅｒｏｎｇ＠１６３．ｃｏｍ；杨　勇（１９７２－），男，博士，教授，研究方向：中医药抗病毒的药效及机制，中医药抗肿瘤的生物疗法探索，通信作者，Ｅ ｍａｉｌ：ｙｙ７２０４＠１６３．ｃｏｍｄｏｉ：１０．１２３６０／ＣＰＢ２０２４０１０１０文献标识码：Ａ文章编号：１００１－１９７８（２０２４）０３－０５９２－０７中国图书分类号：Ｒ２８４ １；Ｒ３２９ ２；Ｒ７３４ ２；Ｒ９７９ １摘要：目的　基于实时无标记细胞分析（ｒｅａｌｔｉｍｅｃｅｌｌｕｌａｒａ ｎａｌｙｓｉｓ，ＲＴＣＡ）技术，以黄芩苷、黄芪甲苷、橙皮苷和顺铂为例进行抗Ａ５４９、Ｈ１２９９肺腺癌活性分析，建立一种反映时间维度变化特征的ＥＣ５０评价新策略。

方法应用ＲＴＣＡＳｏｆｔｗａｒｅＰｒｏ数据分析及ＧｒａｐｈＰａｄＰｒｉｓｍ、ＯｒｉｇｉｎＰｒｏ做图，分别采用终点法和时间维度表征黄芩苷、黄芪甲苷、橙皮苷、顺铂受试液／药的体外抗Ａ５４９、Ｈ１２９９肺腺癌活性。

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20850.01330.806 409 4.50.027060.735 774 3.20.034350.762 15287.90.107180.745 739 5.40.053960.715 227.60.001180.774 105 5.10.007480.647 129 4.90.00780.74475 3.50.002990.639212 3.70.013920.784 201 5.60.009290.599 500 6.30.06138 1.238 >10.00.00042 1.049 290 6.10.019640.625 251 5.60.017330.895 12498.40.075460.68 241 2.20.005560.628 1341 6.70.120620.829 651 6.90.026690.572 224 5.10.010210.672 290 5.30.013220.702 194 1.60.002660.7278>10.00.000620.792 314>10.00.019270.649 32830.001390.381 520 6.30.034550.62153 5.20.004350.904 200 1.30.002460.856 109 3.70.00563143 5.80.013670.825 439 4.20.025470.66 441>10.00.03210.853 17290.80.002020.494 1527.40.009010.741 624 2.40.016810.714 337 6.40.02850.71 173 1.90.002010.614 124 6.30.008010.593 2207.60.010810.602 4708.20.028730.596 221 5.20.00853 1.29 4957.50.036490.714 967.20.003150.52311>10.00.000450.543 184 3.50.006730.588 1064 2.80.017560.666 2437.60.014140.744 257.10.002380.868 105130.022770.574 10130.003550.834 2518.20.017620.63 16669.90.081330.508 1478 4.10.10840.548 14830.008560.8753427.80.015630.587 559.80.002470.6496 1.50.001250.668648.10.002270.561 178 6.20.010850.6 247>10.00.011750.526 202 5.20.013550.701 1484 5.40.069550.546 116 6.40.005740.60572 2.10.00140.58954>10.00.004010.856 351 6.60.011110.57 960 3.90.021660.681 1549.50.006150.597 5790.008180.646 6720.0008353 2.30.00061176 2.60.005320.633 1947.80.016490.569 223 5.60.014960.968 25280.006360.457 1626 5.50.049760.537 1977.10.008730.585 128 5.40.00580.57919>10.00.001080.734 650 3.70.017210.673 9419.10.050830.59113>10.00.00130.598 21090.007690.475 906>10.00.066080.687 350 3.90.005590.396 2247.80.01640.479 1789 5.50.074220.539 419>10.00.014550.491 579 6.80.034280.585 11717.30.084950.73 918.60.003920.471 458.60.00220.655 100 6.20.007370.666 335 5.10.012850.644 964 5.50.038290.59199 5.20.004810.478 302 6.60.012460.48640 2.10.000610.345 392>10.00.021350.719339>10.00.01560.489 5138.90.025150.615 50 4.50.001620.659 1368.50.005580.639 353 5.90.018730.713 495 4.80.011170.321 1147.60.003960.391 156 6.70.022520.706 508 6.50.027410.476 82 5.10.003380.472 410>10.00.020760.421 400>10.00.012770.531 463 4.70.008250.53 728 5.70.012880.253 2367.10.012090.669 180 3.80.004490.45 360>10.00.018270.679 2957.80.035210.653 950 3.40.026580.586 218 5.20.006840.446 2547.50.007820.487。

ＵＨＰＬＣ－ＭＳ/ＭＳ法测定瑞戈非尼中两种痕量基因毒性杂质陈爽ꎬ刘柱ꎬ石云峰ꎬ朱价ꎬ罗英(浙江省食品药品检验研究院ꎬ浙江杭州３１００５８)摘要:目的㊀建立超高效液相色谱－串联质谱(ＵＨＰＬＣ－ＭＳ/ＭＳ)方法测定瑞戈非尼中两类基因毒性杂质[３－氟－４－氨基苯酚(ＳＭ２)㊁３－三氟甲基－４－氯异氰酸苯酯(ＳＭ３)]的含量ꎮ方法㊀采用色谱柱ＡｇｉｌｅｎｔＲＲＨＤＣ１８(２.１ｍｍˑ１００ｍｍꎬ１.８μｍ)ꎬ流动相为０.０５％甲酸水－甲醇ꎬ梯度洗脱ꎬ流速为０.３ｍＬ ｍｉｎ－１ꎬ柱温为３５ħꎻ采用Ａｇｉｌｅｎｔ１２９０－６４７０Ａ液质联用仪和电喷雾离子源(ＡＪＳＥＳＩ)源检测ꎬ正负离子模式采集数据ꎮ结果㊀本方法的专属性㊁线性与范围㊁定量限与检测限㊁准确度㊁精密度及稳定性均符合«中国药典»的验证指标ꎮ结论㊀本方法操作简便ꎬ结果可靠ꎬ用于两批瑞戈非尼中潜在毒性杂质(３－氟－４－氨基苯酚㊁３－三氟甲基－４－氯异氰酸苯酯)的测定ꎮ两批样品中均未检测到ＳＭ２ꎬ而基因毒性杂质ＳＭ３的含量分别为１３.６ˑ１０－６和４１.４ˑ１０－６ꎮ关键词:瑞戈非尼ꎻ基因毒性杂质ꎻ超高效液相色谱－串联质谱中图分类号:Ｒ９２７.１㊀文献标志码:Ａ㊀文章编号:２０９５－５３７５(２０２３)０７－０４８５－００５ｄｏｉ:１０.１３５０６/ｊ.ｃｎｋｉ.ｊｐｒ.２０２３.０７.０１０ＤｅｔｅｒｍｉｎａｔｉｏｎｏｆｔｗｏｔｒａｃｅｇｅｎｏｔｏｘｉｃｉｍｐｕｒｉｔｉｅｓｉｎＲｅｇｏｒａｆｅｎｉｂｂｙＵＨＰＬＣ－ＭＳ/ＭＳＣＨＥＮＳｈｕａｎｇꎬＬＩＵＺｈｕꎬＳＨＩＹｕｎｆｅｎｇꎬＺＨＵＪｉａꎬＬＵＯＹｉｎｇ(ＺｈｅｊｉａｎｇＩｎｓｔｉｔｕｔｅｆｏｒＦｏｏｄａｎｄＤｒｕｇＣｏｎｔｒｏｌꎬＨａｎｇｚｈｏｕ３１００５８ꎬＣｈｉｎａ)Ａｂｓｔｒａｃｔ:Ｏｂｊｅｃｔｉｖｅ㊀ＴｏｅｓｔａｂｌｉｓｈａｎＵＨＰＬＣ－ＭＳ/ＭＳａｎａｌｙｔｉｃａｌｍｅｔｈｏｄｆｏｒｔｈｅｄｅｔｅｒｍｉｎａｔｉｏｎｏｆｇｅｎｏｔｏｘｉｃｉｍｐｕｒｉｔｉｅｓｉｎＲｅｇｏｒａｆｅｎｉｂ.Ｍｅｔｈｏｄｓ㊀ＴｈｅｍｅｔｈｏｄｗａｓａｃｈｉｅｖｅｄｏｎａＡｇｉｌｅｎｔＲＲＨＤＣ１８ｃｏｌｕｍｎ(２.１ｍｍˑ１００ｍｍꎬ１.８μｍ)ｕｔｉｌｉｚｉｎｇａｍｏｂｉｌｅｐｈａｓｅｏｆ０.０５％Ｆｏｒｍｉｃａｃｉｄｗａｔｅｒ(Ａ)－Ｍｅｔｈａｎｏｌ(Ｂ)ｗｉｔｈｇｒａｄｉｅｎｔｅｌｕｔｉｏｎａｔｔｈｅｆｌｏｗｒａｔｅｏｆ０.３ｍＬ ｍｉｎ-１.Ｔｈｅｔｅｍｐｅｒａｔｕｒｅｏｆｃｏｌｕｍｎｗａｓｓｅｔａｔ３５ħ.ＴｈｅＡｇｉｌｅｎｔ１２９０－６４７０ＡＬＣ－ＭＳｗａｓｕｓｅｄｔｏｄｅｔｅｃｔ(ＡＪＳＥＳＩｓｏｕｒｃｅꎬｉｎＭｕｌｔｉｐｌｅＲｅａｃｔｉｏｎＭｏｎｉｔｏｒｉｎｇｍｏｄｅ).Ｒｅｓｕｌｔｓ㊀ＴｈｅｓｐｅｃｉｆｉｃｉｔｙꎬｌｉｎｅａｒｉｔｙａｎｄｒａｎｇｅꎬＬＯＱａｎｄＬＯＤꎬａｃｃｕｒａｃｙꎬｐｒｅｃｉｓｉｏｎａｎｄｓｔａｂｉｌｉｔｙｏｆｔｈｅｍｅｔｈｏｄｗｅｒｅａｌｌｉｎａｃｃｏｒｄａｎｃｅｗｉｔｈｔｈｅｖａｌｉｄａｔｉｏｎｏｆＣｈｉｎｅｓｅＰｈａｒｍａｃｏｐｏｅｉａ.Ｃｏｎｃｌｕｓｉｏｎ㊀Ｔｈｅｐｒｏｐｏｓｅｄｍｅｔｈｏｄｗａｓｓｕｃｃｅｓｓｆｕｌｌｙａｐｐｌｉｅｄｔｏｄｅｔｅｒｍｉｎｅｔｈｅｔｒａｃｅ－ｌｅｖｅｌＰＧＩｓｉｎｔｗｏｂａｔｃｈｅｓｏｆｒｅｇｏｒａｆｅｎｉｂ.ＴｈｅｃｏｎｔｅｎｔｓｏｆＳＭ２ｗｅｒｅｎｏｔｏｂｓｅｒｖｅｄｉｎｔｈｅｔｗｏｂａｔｃｈｅｓｏｆｓａｍｐｌｅｓꎬｗｈｅｒｅａｓｔｈｅｃｏｎｔｅｎｔｓｏｆＳＭ３ｗｅｒｅｄｅｔｅｒｍｉｎｅｄｔｏｂｅ１３.６ˑ１０－６ａｎｄ４１.４ˑ１０－６ꎬｒｅｓｐｅｃｔｉｖｅｌｙ.Ｋｅｙｗｏｒｄｓ:ＲｅｇｏｒａｆｅｎｉｂꎻＧｅｎｏｔｏｘｉｃｉｍｐｕｒｉｔｙꎻＵＨＰＬＣ－ＭＳ/ＭＳ㊀㊀基因毒性杂质(或遗传毒性杂质ꎬｇｅｎｏｔｏｘｉｃｉｍ￣ｐｕｒｉｔｙꎬＧＴＩ)是少量即能引起细胞ＤＮＡ损伤ꎬ诱导基因突变ꎬ并具有致癌倾向的化合物[１]ꎮ欧洲药品管理局(ＥＭＥＡ)㊁美国食品药品管理局(ＦＤＡ)及人用药品注册技术要求国际协调会(ＩＣＨ)先后颁布了基因毒性杂质控制的指导文件[２－４]ꎬ推荐以毒理学关注阈值(ＴＴＣꎬ１.５μｇ ｄ－１)来控制用药风险ꎬ并发布基因毒性警示结构基团ꎮ瑞戈非尼(Ｒｅｇｏｒａｆｅｎｉｂ)是拜耳药业研发并于２０１７年３月在国内上市的一种新型口服多靶点的磷酸激酶抑制剂ꎬ本品以促进肿瘤血管生成和细胞增殖的多种蛋白激酶为靶标ꎬ达到治疗晚期及转移性结肠癌㊁进展期肝细胞癌㊁胃肠道恶性间质肿瘤㊁儿童神经母细胞瘤和胶质母细胞瘤及一些转移性实体肿瘤的目的ꎮ瑞戈非尼的合成路线中起始物料ＳＭ２㊁ＳＭ３结构中均含有苯胺类基因毒性警示结构基团ꎬ具有潜在基因毒性ꎬ详见图１ꎮ瑞戈非尼已收载于«中国药典»和«欧洲药典»ꎬ已经报道了一种ＬＣ－ＭＳ/ＭＳ法定量同时测定肝细胞癌患者血浆中瑞戈非尼中及其两种活性代谢物的含量的方法[１７]ꎬ但目前也未见相关文献报道本品中两种苯胺类基因毒性杂质ＳＭ２㊁ＳＭ３的测定方法ꎮ㊀作者简介:陈爽ꎬ女ꎬ硕士ꎬ副主任药师ꎬ研究方向:药物质量研究㊁标准提高研究ꎬＥ－ｍａｉｌ:ｃｒｅｄｅａｒ＠１２６.ｃｏｍＡ.３－氟－４－氨基苯酚(ＳＭ２)ꎻＢ.３－三氟甲基－４－氯异氰酸苯酯(ＳＭ３)图１㊀瑞戈非尼中苯胺类基因毒性杂质结构因此建立超高效液相色谱－串联质谱(ＵＨＰＬＣ－ＭＳ/ＭＳ)方法对瑞戈非尼中少量的苯胺类基因毒性杂质的含量进行控制ꎮ本研究依照ＩＣＨ推荐的条件ꎬ建立了方法并从方法的专属性㊁溶液稳定性㊁线性与范围㊁定量限与检测限㊁准确度㊁精密度方面进行了方法学研究ꎬ本法灵敏㊁可靠㊁简便ꎬ为瑞戈非尼工艺过程控制和质量保障提供参考依据ꎮ１㊀仪器与试剂Ａｇｉｌｅｎｔ１２９０－６４７０ＡＬＣ－ＭＳ联用系统(美国安捷伦)ꎬ配备电喷雾离子源(ＡＪＳＥＳＩ)ꎬ质谱软件为ＡｇｉｌｅｎｔＭａｓｓＨｕｎｔｅｒ工作站ꎻＸＳ２０５ＤＵ百万分之一电子天平(瑞士梅特勒托利多公司)ꎮＡｇｉｌｅｎｔＲＲＨＤＣ１８(２.１ｍｍˑ１００ｍｍꎬ１.８μｍꎬ填料:十八烷基硅烷键合硅胶ꎻＡｇｉｌｅｎｔ公司)ꎮ３－氟－４－氨基苯酚对照品(批号:Ｐ１１５６８１１ꎬ纯度:９８％)ꎬ３－三氟甲基－４－氯异氰酸苯酯对照品(杭州华东医药集团新药研究院有限公司ꎬ批号:１９１２０１ꎬ纯度:９９.７６％)ꎻ瑞戈非尼(批号:ＲＧＦＮ－Ｂ－１９０９０１㊁Ｚ１９３－Ａ１９１１００１ꎬ由Ａ公司提供)ꎻ甲醇(默克试剂有限公司)㊁异丙醇㊁甲酸(阿拉丁有限公司)均为色谱级ꎮ２㊀方法与结果２.１㊀色谱条件㊀液相色谱柱:ＡｇｉｌｅｎｔＲＲＨＤＣ１８(２.１ｍｍˑ１００ｍｍꎬ１.８μｍ)ꎻ流速０.３ｍＬ ｍｉｎ－１ꎻ０.０５％甲酸水为流动相Ａꎬ甲醇为流动相Ｂꎬ梯度洗脱(０~１.２ｍｉｎꎬ１０％Ｂꎻ１.２~４.０ｍｉｎꎬ１０％Ｂң６０％Ｂꎻ４.０~８.０ｍｉｎꎬ６０％Ｂꎻ８.０~１３.０ｍｉｎꎬ９５％Ｂꎻ１３.０~１３.１０ｍｉｎꎬ９５％Ｂң１０％Ｂꎻ１３.１０~１５.０ｍｉｎꎬ１０％Ｂ)ꎻ柱温３５ħꎻ进样体积５μＬꎮ取０.６ｎｇ ｍＬ－１混标对照品溶液ꎬ在ＭＲＭ模式下进样分析ꎬ结果如图２所示ꎬ各杂质之间分离度良好ꎮ２.２㊀质谱条件㊀采用电喷雾离子源(ＡＪＳＥＳＩ)ꎬ正负离子检测模式ＭＲＭ采集(ＳＭ２正离子模式㊁ＳＭ３㊀ＳＭ２[３－氟－４－氨基苯酚(３－Ｆｌｕｏｒｏ－４－ａｍｉｎｏｐｈｅｎｏｌ)]ꎻＳＭ３[３－三氟甲基－４－氯异氰酸苯酯(３－ｔｒｉｆｌｕｏｒｏｍｅｔｈｙｌ－４－ｐｈｅｎｙｌ￣ｃｈｌｏｒｏｉｓｏｃｙａｎａｔｅ)]图２㊀混合对照品溶液总离子流图负离子模式)ꎬ参数见表１ꎮ喷雾电压为３.５ｋＶꎬ鞘气为２５０ħꎬ鞘气流速为１１Ｌ ｍｉｎ－１ꎬ雾化气为４５ｐｓｉꎬ干燥气温度为３００ħꎬ干燥气流速为５Ｌ ｍｉｎ－１ꎮ母离子及子离子:取０.６μｇ ｍＬ－１混标对照品溶液ꎬ在Ｓｃａｎ模式下进样ꎬ进行ＭＳ分析ꎬ两种基因毒性杂质的质谱图如图４所示ꎬ其中３－氟－４－氨基苯酚的母离子为ｍ/ｚ１２８.０５ꎬ３－三氟甲基－４－氯异氰酸苯酯的母离子为ｍ/ｚ２８０.００ꎮ各基因毒性杂质的ＭＲＭ参数见表１ꎬ定量离子及定性离子的选择见表１ꎮ表１㊀ＭＲＭ采集参数设置化合物类型母离子(ｍ/ｚ)子离子(ｍ/ｚ)碰撞能量/ｅＶ碎裂电压/Ｖ扫描时间/ｍｓＳＭ２(ＥＳＩ＋)ＳＭ３(ＥＳＩ－)定量离子１２８.０５１０８１７定性离子１２８.０５８１.１２５定量离子２８０２１９.９１３定性离子２８０１９９.９２５８５１１２１００㊀ＳＭ２[３－氟－４－氨基苯酚(３－ｆｌｕｏｒｏ－４－ａｍｉｎｏｐｈｅｎｏｌ)]ꎻＳＭ３[３－三氟甲基－４－氯异氰酸苯酯(３－ｔｒｉｆｌｕｏｒｏｍｅｔｈｙｌ－４－ｐｈｅｎｙｌ￣ｃｈｌｏｒｏｉｓｏｃｙａｎａｔｅ)]图３㊀两种基因毒性杂质的质谱图２.３㊀混标对照品溶液㊀精密称取３－氟－４－氨基苯酚对照品㊁３－三氟甲基－４－氯异氰酸苯酯对照品各约６ｍｇꎬ加稀释剂异丙醇溶解并稀释成６ｎｇ ｍＬ－１的混标对照品储备液ꎮ精密量取适量ꎬ稀释１０倍ꎬ作为０.６ｎｇ ｍＬ－１混标对照品溶液ꎮ２.４㊀供试品溶液㊀取本品约２０ｍｇꎬ精密称定ꎬ置２０ｍＬ容量瓶ꎬ用稀释剂溶解并稀释到刻度ꎬ摇匀ꎻ再精密量取０.１ｍＬꎬ置１０ｍＬ容量瓶ꎮ异丙醇稀释至刻度ꎬ摇匀ꎬ作为供试品溶液(１０μｇ ｍＬ－１)ꎮ２.５㊀１００％加标供试品溶液㊀取本品约２０ｍｇꎬ精密称定ꎬ置２０ｍＬ量瓶中ꎬ用异丙醇溶解稀释至刻度ꎬ摇匀ꎻ再精密量取０.１ｍＬꎬ置１０ｍＬ容量瓶ꎬ加入１ｍＬ对照品储备液后ꎬ再用异丙醇稀释至刻度ꎬ摇匀ꎬ作为１００％加标供试品溶液ꎮ２.６㊀方法学考察㊀２.６.１㊀专属性试验㊀取稀释剂照上述条件进行试验ꎬ记录色谱图ꎬ结果显示稀释剂不干扰测定ꎬ表明该方法专属性良好ꎮ２.６.２㊀线性关系㊁检测限和定量限㊀精密量取混标对照品储备液适量ꎬ加稀释剂稀释得到浓度分别为０.１５㊁０.３㊁０.４５㊁０.６㊁０.９㊁１.２ｎｇ ｍＬ－１的混标对照品溶液ꎬ进样分析ꎬ绘制标准曲线ꎬ得线性方程(见表２)ꎮ逐步稀释ꎬ考察定量限及检测限(见图４)ꎮ表２㊀瑞戈非尼中两种基因毒性杂质的线性范围㊁定量检测限及精密度参数ＳＭ２ＳＭ３线性方程Ｙ＝４７９９.５Ｘ＋２７.１Ｙ＝８９３.３Ｘ＋２７.１范围/ｎｇ ｍＬ－１０.１５~１.１７０.１５~１.１９ｒ０.９９９９０.９９９１检测限/ｎｇ ｍＬ－１０.０６０.０６定量限/ｎｇ ｍＬ－１０.１５０.１５精密度ＲＳＤ(％)１.３２１.９７中间精密度ＲＳＤ(％)２.５３２.２４图４㊀定量限和检测限ＭＲＭ质谱图２.６.３㊀稳定性试验㊀取 ２.３ 项下０.６ｎｇ ｍＬ－１的混标对照品溶液ꎬ ２.５ 项下１０μｇ ｍＬ－１的１００％加标供试品溶液ꎬ连续进样１６ｈ进行分析ꎬ各峰面积偏离度的绝对值在８ｈ内均小于１７.２２％ꎬ表明对照品溶液和１００％加标供试品溶液在８ｈ内稳定性良好ꎮ２.６.４㊀准确度和精密度试验㊀精密称取瑞戈非尼(批号:ＲＧＦＮ－Ｂ－１９０９０１㊁Ｚ１９３－Ａ１９１１００１)约２０ｍｇꎬ一式１２份ꎬ分别置２０ｍＬ量瓶中ꎬ用异丙醇溶解稀释至刻度ꎬ摇匀ꎻ再精密量取０.１ｍＬꎬ置１０ｍＬ容量瓶ꎬ精密加入 ２.３ 项下浓度为６ｎｇ ｍＬ－１的混标对照品储备液溶液０.２５㊁０.５㊁１.０㊁１.５ｍＬꎬ分别溶解并稀释至刻度ꎬ摇匀ꎬ即得低(０.１５ｎｇ ｍＬ－１ꎬ３份)㊁中(０.３０ｎｇ ｍＬ－１ꎬ３份)㊁高(０.６０ｎｇ ｍＬ－１ꎬ３份)㊁极高(０.９０ｎｇ ｍＬ－１ꎬ３份)４个浓度的供试品加标溶液ꎬ取混标对照品溶液和各加标供试品溶液分别进样分析ꎬ外标法计算回收率结果见表３ꎬ各基因毒性杂质回收率均在９０.６４％~１０５.４４％之间ꎬＲＳＤ均小于３.５０％ꎬ表明各基因毒性杂质回收率和精密度良好ꎮ２.７㊀样品测定㊀取瑞戈非尼供试品２批(批号:ＲＧＦＮ－Ｂ－１９０９０１㊁Ｚ１９３－Ａ１９１１００１)ꎬ每批精密称取２份约２０ｍｇꎬ置２０ｍＬ量瓶中ꎬ加稀释剂溶解并稀释至刻度ꎬ摇匀ꎻ再精密量取０.１ｍＬꎬ置１０ｍＬ容量瓶ꎬ异丙醇稀释至刻度ꎬ摇匀ꎬ作为供试品溶液(１０μｇ ｍＬ－１)ꎬ进样分析ꎬ结果２批次瑞戈非尼中ＳＭ２均未检出ꎬＳＭ３均有检出(见表４)ꎮ表３㊀瑞戈非尼中各基因毒性杂质的回收率化合物加入量测得量回收率平均回收ＲＳＤ(％)ＳＭ３１.５０２.９９５.９８８.９８２.７７１０１.３３２.７８１００.６７３.９７９０.６４４.１３９５.９９４.３２１０２.３４７.０５９６.８２７.２２９９.６７７.４２１０２.８４１０.２５１００.００１０.０９９８.３３１０.２３９９.８９９９.１５３.４１表４㊀瑞戈非尼中基因毒性杂质检测结果批号ＳＭ２含量(％)ＳＭ３含量(％)Ｚ１９３－Ａ１９１１００１０.０００００.００１３６ＲＧＦＮ－Ｂ－１９０９０１０.０００００.００４１４３㊀讨论３.１㊀瑞戈非尼的合成工艺㊀如图１所示ꎬ以３－氟－４－氨基苯酚和Ｎ－甲基－４－氯－２－吡啶甲酰胺为原料㊁氢氧化钠为碱ꎬＴＨＦ为溶剂ꎬ回流反应得到中间体４－(４－氨基－３－氟苯氧基)－Ｎ－甲基吡啶－２－甲酰胺ꎬ该中间体再与４－氯－３－三氟甲基苯胺以羰基连接得到瑞戈非尼无水物(４－[４－[[[４－氯－３－(三氟甲基)苯基]氨基甲酰]氨基]－３－氟苯氧基]－Ｎ－甲基吡啶－２－甲酰胺)ꎬ加水而成ꎮ合成路线中涉及多个起始物料及中间体带有基因警示结构ꎬ因此ꎬ必须对瑞戈非尼合成路线中引入的带有基因警示结构的潜在基因毒性杂质进行控制ꎮ３.２㊀控制限度㊀参照瑞戈非尼片药品使用说明书ꎬ建议每日最低剂量为８０ｍｇꎬ每日最高剂量为１６０ｍｇꎮ因为瑞戈非尼为抗肿瘤用药ꎬ可以不以ＩＣＨＭ７推荐的最严格的毒理学关注阈值(ＴＴＣꎬ１.５μｇ ｄ－１)为控制限制ꎮ本研究基于治疗期ꎬ以相对严格的１~１０年治疗期的ＴＴＣ(１０μｇ ｄ－１)为控制限制计算ꎬ瑞戈非尼中基因毒性杂质限度应为６２.５ˑ１０－６ꎬ本研究以６０ˑ１０－６作为瑞戈非尼中基因毒性杂质的控制限度ꎮ本次研究用样品中未检出ＳＭ２ꎬ检出的ＳＭ３分别为１３.６ˑ１０－６㊁４１.４ˑ１０－６ꎬ均符合限度要求ꎬ但ＳＭ３检出量已超３０％限度ꎬ值得关切ꎮ３.３㊀色谱条件的优化㊀色谱条件优化试验中ꎬ探索了甲醇㊁异丙醇和乙腈３种有机相(Ｂ相)ꎬ水相(Ａ相)中添加不同比率的酸(０.１％甲酸水ꎬ０.０５％甲酸水和０.１％乙酸水)对分离度和灵敏度的影响ꎮ结果显示ꎬ采用ＡｇｉｌｅｎｔＲＲＨＤＣ１８色谱柱ꎬ当流动相０.０５％甲酸水溶液－甲醇梯度洗脱时ꎬ３－氟－４－氨基苯酚(ＳＭ２)㊁３－三氟甲基－４－氯异氰酸苯酯(ＳＭ３)响应较好ꎬ且两个化合物分离度良好ꎬ故流动相中选择０.０５％甲酸水ꎮ本试验考察了流速０.２~０.４ｍＬ ｍｉｎ－１范围内对苯胺类物质响应和峰形的影响ꎬ发现流速０.３ｍＬ ｍｉｎ－１时ꎬＳＭ２ꎬＳＭ３离子化效率最高ꎬ出峰时间合适ꎬ且峰形较好ꎮ３.４㊀质谱解析㊀３－氟－４－氨基苯酚的质谱图中ｍ/ｚ１２８.０５为[Ｍ＋Ｈ]＋离子ꎬ主要的碎片离子有ｍ/ｚ１０８和ｍ/ｚ８１.１ꎬ其中ｍ/ｚ１０８为去氟基产物ꎬ反映了３－氟－４－氨基苯酚的结构特征ꎮ试验对３－氟－４－氨基苯酚２个离子对ｍ/ｚ１２８.０５ң１０８和ｍ/ｚ１２８.０５ң８１.１的ＭＲＭ参数见表１ꎬ其中ꎬ离子对ｍ/ｚ１２８.０５ң１０８的信噪比较离子对ｍ/ｚ１２８.０５ң８１.１更高ꎬ因此选择ｍ/ｚ１２８.０５ң１０８作为定量离子对ꎬｍ/ｚ１２８.０５ң８１.１作为定性离子对ꎮ本法中ＳＭ３检测采用异丙醇衍生化ꎬ分析对象为ＳＭ３与异丙醇衍生后的产物ꎮ衍生产物精确分子量为２８１.０４ꎬ其[Ｍ－Ｈ]－为２８０ꎬ反应过程见图５ꎮ图５㊀ＳＭ３衍生化反应(下转第５２５页)ｆａｔｔｙａｃｉｄｂｉｏｇｅｎｅｓｉｓ:ａｎｅｗｆａｍｉｌｙｏｆａｎｔｉ－ｃａｎｃｅｒａｇｅｎｔｓ?[Ｊ].ＣｕｒｒＰｈａｒｍＢｉｏｔｅｃｈｎｏｌꎬ２００６ꎬ７(６):４８３－４９３.[８７]ＣＨＥＮＸꎬＨＵＡＮＧＫꎬＨＵＳꎬｅｔａｌ.ＦＡＳＮ－ＭｅｄｉａｔｅｄＬｉｐｉｄＭｅｔａｂｏｌｉｓｍＲｅｇｕｌａｔｅｓＧｏｏｓｅＧｒａｎｕｌｏｓａＣｅｌｌｓＡｐｏｐｔｏｓｉｓａｎｄＳｔｅｒｏｉｄｏｇｅｎｅｓｉｓ[Ｊ].ＦｒｏｎｔｉｎＰｈｙｓｉｏｌꎬ２０２０(１１):６００.[８８]ＣＨＯＹＫꎬＳＯＮＹꎬＫＩＭＳＮꎬｅｔａｌ.ＭｉｃｒｏＲＮＡ－１０ａ－５ｐｒｅｇｕｌａｔｅｓｍａｃｒｏｐｈａｇｅｐｏｌａｒｉｚａｔｉｏｎａｎｄｐｒｏｍｏｔｅｓｔｈｅｒａｐｅｕｔｉｃａｄｉｐｏｓｅｔｉｓｓｕｅｒｅｍｏｄｅｌｉｎｇ[Ｊ].ＭｏｌＭｅｔａｂꎬ２０１９(２９):８６－９８.[８９]ＸＵＥＭꎬＹＡＮＧＭｘｉｎｇꎬＺＨＡＮＧＷꎬｅｔａｌ.Ｃｈａｒａｃｔｅｒｉｚａｔｉｏｎꎬｐｈａｒｍａｃｏｋｉｎｅｔｉｃｓꎬａｎｄｈｙｐｏｇｌｙｃｅｍｉｃｅｆｆｅｃｔｏｆｂｅｒｂｅｒｉｎｅｌｏａｄｅｄｓｏｌｉｄｌｉｐｉｄｎａｎｏｐａｒｔｉｃｌｅｓ[Ｊ].ＩｎｔＪＮａｎｏｍｅｄｉｃｉｎｅꎬ２０１３(８):４６７７－４６８７.[９０]ＸＵＥＭꎬＺＨＡＮＧＬꎬＹＡＮＧＭＸꎬｅｔａｌ.Ｂｅｒｂｅｒｉｎｅ－ｌｏａｄｅｄｓｏｌｉｄｌｉｐｉｄｎａｎｏｐａｒｔｉｃｌｅｓａｒｅｃｏｎｃｅｎｔｒａｔｅｄｉｎｔｈｅｌｉｖｅｒａｎｄａｍｅｌｉｏｒａｔｅｈｅｐａｔｏｓｔｅａｔｏｓｉｓｉｎｄｂ/ｄｂｍｉｃｅ[Ｊ].ＩｎｔＪＮａｎｏ￣ｍｅｄｉｃｉｎｅꎬ２０１５(１０):５０４９－５０５７.[９１]ＺＨＡＯＪꎬＺＨＡＯＱꎬＬＵＪＺꎬｅｔａｌ.ＮａｔｕｒａｌＮａｎｏ－ＤｒｕｇＤｅ￣ｌｉｖｅｒｙＳｙｓｔｅｍｉｎＣｏｐｔｉｄｉｓＲｈｉｚｏｍａＥｘｔｒａｃｔｗｉｔｈＭｏｄｉｆｉｅｄＢｅｒｂｅｒｉｎｅＨｙｄｒｏｃｈｌｏｒｉｄｅＰｈａｒｍａｃｏｋｉｎｅｔｉｃｓ[Ｊ].ＩｎｔＪＮａｎｏｍｅｄｉｃｉｎｅꎬ２０２１(１６):６２９７－６３１１.[９２]ＤＵＯＮＧＴＴꎬＩＳＯＭÄＫＩＡꎬＰＡＡＶＥＲＵꎬｅｔａｌ.Ｎａｎｏｆｏｒｍｕ￣ｌａｔｉｏｎａｎｄＥｖａｌｕａｔｉｏｎｏｆＯｒａｌＢｅｒｂｅｒｉｎｅ－ＬｏａｄｅｄＬｉｐｏｓｏｍｅｓ[Ｊ].Ｍｏｌｅｃｕｌｅｓꎬ２０２１ꎬ２６(９):２５９１.[９３]ＳＡＨＩＢＺＡＤＡＭＵＫꎬＺＡＨＯＯＲＭꎬＳＡＤＩＱＡꎬｅｔａｌ.Ｂｉｏ￣ａｖａｉｌａｂｉｌｉｔｙａｎｄｈｅｐａｔｏｐｒｏｔｅｃｔｉｏｎｅｎｈａｎｃｅｍｅｎｔｏｆｂｅｒｂｅｒｉｎｅａｎｄｉｔｓｎａｎｏｐａｒｔｉｃｌｅｓｐｒｅｐａｒｅｄｂｙｌｉｑｕｉｄａｎｔｉｓｏｌ￣ｖｅｎｔｍｅｔｈｏｄ[Ｊ].ＳａｕｄｉＪＢｉｏｌＳｃｉꎬ２０２１ꎬ２８(１):３２７－３３２.[９４]ＣＨＯＷＹＬꎬＳＯＧＡＭＥＭꎬＳＡＴＯＦ.１３－Ｍｅｔｈｙｌｂｅｒｂｅｒｉｎｅꎬａｂｅｒｂｅｒｉｎｅａｎａｌｏｇｕｅｗｉｔｈｓｔｒｏｎｇｅｒａｎｔｉ－ａｄｉｐｏｇｅｎｉｃｅｆｆｅｃｔｓｏｎｍｏｕｓｅ３Ｔ３－Ｌ１ｃｅｌｌｓ[Ｊ].ＳｃｉｅｎｔｉｆｉｃＲｅｐｏｒｔｓꎬ２０１６ꎬ６(１):３８１２９.[９５]ＲＥＢＥＬＬＯＣＪꎬＧＲＥＥＮＷＡＹＦＬ.Ｏｂｅｓｉｔｙｍｅｄｉｃａｔｉｏｎｓｉｎｄｅｖｅｌｏｐｍｅｎｔ[Ｊ].ＥｘｐｅｒｔＯｐｉｎＩｎｖｅｓｔｉｇＤｒｕｇｓꎬ２０２０ꎬ２９(１):６３－７１.(收稿日期:２０２２－１２－３１)(上接第４８８页)㊀㊀ＳＭ３衍生产物ꎬ在质谱离子源中ꎬ子离子为酯键断裂后重排的ｍ/ｚꎬ碎裂途径见图６ꎮ图６㊀ＳＭ３子离子裂解途径本研究建立了超高效液相色谱－串联质谱法测定瑞戈非尼中的２种基因毒性杂质(３－氟－４－氨基苯酚(ＳＭ２)㊁３－三氟甲基－４－氯异氰酸苯酯(ＳＭ３)ꎬ并进行了方法学验证ꎬ可以有效控制瑞戈非尼的潜在基因毒性杂质ꎮ该方法专属性强ꎬ灵敏度高ꎬ满足基因毒性杂质测定要求ꎬ可以作为瑞戈非尼原料药中基因毒性杂质的质控方法ꎮ参考文献:[１]㊀ＥｕｒｏｐｅａｎＭｅｄｉｃｉｎｅｓＡｇｅｎｃｙ(ＥＭＡ).ＧｕｉｄｅｌｉｎｅｏｎｔｈｅＬｉｍｉｔｓｏｆＧｅｎｏｔｏｘｉｃＩｍｐｕｒｉｔｉｅｓ(欧洲药品管理局关于基因毒性杂质限度指导原则)[ＥＢ/ＯＬ].[２０１５－０３－０２].ｈｔｔｐ://ｗｗｗ.Ｅｍａ.Ｅｕｒｏｐａ.ｅｕ/ｄｏｃｓ/ｅｎ＿ＧＢ/ｄｏｃｕｍｅｎｔ＿ｌｉ￣ｂｒａｒｙ/Ｓｃｉｅｎｔｉｆｉｃ＿ｇｕｉｄｅｌｉｎｅ/２００９/０９/ＷＣ５００００２９０３.ｐｄｆ.[２]ＥｕｒｏｐｅａｎＭｅｄｉｃｉｎｅｓＡｇｅｎｃｙ(ＥＭＡ).ＱｕｅｓｔｉｏｎｓａｎｄＡｎｓｗｅｒｓｏｎｔｈｅＧｕｉｄｅｌｉｎｅｏｎｔｈｅＬｉｍｉｔｓｏｆＧｅｎｏｔｏｘｉｃＩｍ￣ｐｕｒｉｔｉｅｓ[ＥＢ/ＯＬ].(２０１０－０９－２３)[２０１５－０３－０２].ｈｔｔｐ//ｗｗｗ.ｅｍａ.ｅｕｒｏｐａ.ｅｕ/ｄｏｃｓ/ｅｎ＿ＧＢ/ｄｏｃｕｍｅｎｔ＿ｌｉｂｒａｒｙ/Ｓｃｉ￣ｅｎｔｉｆｉｃ＿ｇｕｉｄｅｌｉｎｅ/２００９/０９/ＷＣ５００００２９０７.ｐｄｆ.[３]ＦｏｏｄａｎｄＤｒｕｇＡｄｍｉｎｉｓｔｒａｔｉｏｎ(ＦＤＡ).ＧｅｎｏｔｏｘｉｃａｎｄＣａｒ￣ｃｉｎｏｇｅｎｉｃＩｍｐｕｒｉｔｉｅｓＩｎＤｒｕｇＳｕｂｓｔａｎｃｅｓａｎｄＰｒｏｄｕｃｔｓ:ＲｅｃｏｍｍｅｎｄｅｄＡｐｐｒｏａｃｈｅｓ.[２０１５－０８－２６].[４]ＩｎｔｅｒｎａｔｉｏｎａｌＣｏｎｆｅｒｅｎｃｅｏｎＨａｒｍｏｎｉｚａｔｉｏｎｏｆＴｅｃｈｎｉｃａｌＲｅｑｕｉｒｅｍｅｎｔｓｆｏｒＲｅｇｉｓｔｒａｔｉｏｎｏｆＰｈａｒｍａｃｅｕｔｉｃａｌｓｆｏｒＨｕ￣ｍａｎＵｓｅ(ＩＣＨ).ＡｓｓｅｓｓｍｅｎｔａｎｄＣｏｎｔｒｏｌｏｆＤＮＡＲｅａｃｔｉｖｅ(Ｍｕｔａｇｅｎｉｃ)ＩｍｐｕｒｉｔｉｅｓｉｎＰｈａｒｍａｃｅｕｔｉｃａｌｓｔｏＬｉｍｉｔＰｏ￣ｔｅｎｔｉａｌＣａｒｃｉｎｏｇｅｎｉｃＲｉｓｋＭ７(Ｒ１).[２０１５－０８－２６].[５]ＲＡＭＡＮＮＶＶＳＳꎬＰＲＡＳＡＤＡＶＳＳꎬＲＥＤＤＹＫＲ.Ｓｔｒａｔｅｇｉｅｓｆｏｒｔｈｅｉｄｅｎｔｉｆｉｃａｔｉｏｎꎬｃｏｎｔｒｏｌａｎｄｄｅｔｅｒｍｉｎａｔｉｏｎｏｆｇｅｎｏｔｏｘｉｃｉｍｐｕｒｉｔｉｅｓｉｎｄｒｕｇｓｕｂｓｔａｎｃｅｓ:ａｐｈａｒｍａｃｅｕｔｉｃａｌｉｎｄｕｓｔｒｙｐｅｒｓｐｅｃｔｉｖｅｓ[Ｊ].ＪＰｈａｒｍＢｉｏｍｅｄＡｎａｌꎬ２０１１ꎬ５５(４):６６２－６６７.[６]ＳＺＥＫＥＬＹＧꎬＡＭＯＲＥＳＤＥＳＯＵＳＡＭＣꎬＧＩＬＭꎬｅｔａｌ.ＧｅｎｏｔｏｘｉｃＩｍｐｕｒｉｔｉｅｓｉｎＰｈａｒｍａｃｅｕｔｉｃａｌＭａｎｕｆａｃｔｕｒｉｎｇ:ＳｏｕｒｃｅｓꎬＲｅｇｕｌａｔｉｏｎｓꎬａｎｄＭｉｔｉｇａｔｉｏｎ[Ｊ].ＣｈｅｍｉｃａｌＲｅｖｉｅｗｓꎬ２０１５ꎬ１１５(１６):８１８２－８２２９.[７]甘勇军ꎬ王以武ꎬ张量ꎬ等.瑞戈非尼的合成工艺优化[Ｊ].中国医药工业杂志ꎬ２０２０ꎬ５１(２):１９６－１９９.[８]曹琳ꎬ罗淑青ꎬ章燕ꎬ等.ＬＣ－ＱＱＱ－ＭＳ/ＭＳ分析苯磺酸氨氯地平中痕量苯磺酸酯类基因毒性杂质[Ｊ].中国现代应用药学ꎬ２０２０ꎬ３７(１１):１２９６－１３００.[９]芦飞ꎬ邢珊珊ꎬ王咏.ＬＣ－ＭＳ测定醋酸甲羟孕酮中对甲苯磺酸甲酯及对甲苯磺酸乙酯的含量[Ｊ].中国现代应用药学ꎬ２０１８ꎬ３５(５):６５７－６５９.(收稿日期:２０２３－０２－２４)。